Product datasheet
Anti-Amyloid Precursor Protein antibody ab15272
22 Abreviews 10 References 6 Images
Overview
Product name
Anti-Amyloid Precursor Protein antibody
Description
Rabbit polyclonal to Amyloid Precursor Protein
Specificity
Ab15272 recognizes APP (beta-APP770) and is predicted to recognize beta-APP695 and betaAPP751. The antibody stains both cytoplasmic and extracellular areas.
Tested applications
IP, ICC/IF, ICC, IHC-FoFr, WB, IHC-P
Species reactivity
Reacts with: Mouse, Rat, Human
Immunogen
Synthetic peptide corresponding to Human Amyloid Precursor Protein aa 44-62 (N terminal).
Sequence:
HMNVQNGKWDSDPSGTKTC
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Positive control
Run BLAST with
Brain
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer
pH: 7.40
Preservative: 0.1% Sodium azide
Constituents: 0.0268% PBS, BSA
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab15272 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Abreviews
Notes
IP
Use at an assay dependent concentration. PubMed: 21163940
ICC/IF
1/100.
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Application
Abreviews
Notes
ICC
Use at an assay dependent concentration.
IHC-FoFr
1/100.
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 87 kDa. ab15272
reacts strongly with some immature isoforms and cleavage products (Range 3070 kDa). Weaker signal from mature isoforms (over 100 kDa) (this maybe due to
steric inhibition by post-translational modifications).
IHC-P
1/100.
Target
Function
Functions as a cell surface receptor and performs physiological functions on the surface of
neurons relevant to neurite growth, neuronal adhesion and axonogenesis. Involved in cell mobility
and transcription regulation through protein-protein interactions. Can promote transcription
activation through binding to APBB1-KAT5 and inhibits Notch signaling through interaction with
Numb. Couples to apoptosis-inducing pathways such as those mediated by G(O) and JIP.
Inhibits G(o) alpha ATPase activity (By similarity). Acts as a kinesin I membrane receptor,
mediating the axonal transport of beta-secretase and presenilin 1. Involved in copper
homeostasis/oxidative stress through copper ion reduction. In vitro, copper-metallated APP
induces neuronal death directly or is potentiated through Cu(2+)-mediated low-density
lipoprotein oxidation. Can regulate neurite outgrowth through binding to components of the
extracellular matrix such as heparin and collagen I and IV. The splice isoforms that contain the
BPTI domain possess protease inhibitor activity. Induces a AGER-dependent pathway that
involves activation of p38 MAPK, resulting in internalization of amyloid-beta peptide and leading
to mitochondrial dysfunction in cultured cortical neurons. Provides Cu(2+) ions for GPC1 which
are required for release of nitric oxide (NO) and subsequent degradation of the heparan sulfate
chains on GPC1.
Beta-amyloid peptides are lipophilic metal chelators with metal-reducing activity. Bind transient
metals such as copper, zinc and iron. In vitro, can reduce Cu(2+) and Fe(3+) to Cu(+) and
Fe(2+), respectively. Beta-amyloid 42 is a more effective reductant than beta-amyloid 40. Betaamyloid peptides bind to lipoproteins and apolipoproteins E and J in the CSF and to HDL
particles in plasma, inhibiting metal-catalyzed oxidation of lipoproteins. Beta-APP42 may
activate mononuclear phagocytes in the brain and elicit inflammatory responses. Promotes both
tau aggregation and TPK II-mediated phosphorylation. Interaction with overexpressed HADH2
leads to oxidative stress and neurotoxicity. Also binds GPC1 in lipid rafts.
Appicans elicit adhesion of neural cells to the extracellular matrix and may regulate neurite
outgrowth in the brain.
The gamma-CTF peptides as well as the caspase-cleaved peptides, including C31, are potent
enhancers of neuronal apoptosis.
N-APP binds TNFRSF21 triggering caspase activation and degeneration of both neuronal cell
bodies (via caspase-3) and axons (via caspase-6).
Tissue specificity
Expressed in all fetal tissues examined with highest levels in brain, kidney, heart and spleen.
Weak expression in liver. In adult brain, highest expression found in the frontal lobe of the cortex
and in the anterior perisylvian cortex-opercular gyri. Moderate expression in the cerebellar
cortex, the posterior perisylvian cortex-opercular gyri and the temporal associated cortex. Weak
expression found in the striate, extra-striate and motor cortices. Expressed in cerebrospinal
fluid, and plasma. Isoform APP695 is the predominant form in neuronal tissue, isoform APP751
and isoform APP770 are widely expressed in non-neuronal cells. Isoform APP751 is the most
abundant form in T-lymphocytes. Appican is expressed in astrocytes.
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Involvement in disease
Alzheimer disease 1
Cerebral amyloid angiopathy, APP-related
Sequence similarities
Belongs to the APP family.
Contains 1 BPTI/Kunitz inhibitor domain.
Domain
The basolateral sorting signal (BaSS) is required for sorting of membrane proteins to the
basolateral surface of epithelial cells.
The NPXY sequence motif found in many tyrosine-phosphorylated proteins is required for the
specific binding of the PID domain. However, additional amino acids either N- or C-terminal to
the NPXY motif are often required for complete interaction. The PID domain-containing proteins
which bind APP require the YENPTY motif for full interaction. These interactions are independent
of phosphorylation on the terminal tyrosine residue. The NPXY site is also involved in clathrinmediated endocytosis.
Post-translational
modifications
Proteolytically processed under normal cellular conditions. Cleavage either by alpha-secretase,
beta-secretase or theta-secretase leads to generation and extracellular release of soluble APP
peptides, S-APP-alpha and S-APP-beta, and the retention of corresponding membraneanchored C-terminal fragments, C80, C83 and C99. Subsequent processing of C80 and C83 by
gamma-secretase yields P3 peptides. This is the major secretory pathway and is nonamyloidogenic. Alternatively, presenilin/nicastrin-mediated gamma-secretase processing of
C99 releases the amyloid beta proteins, amyloid-beta 40 (Abeta40) and amyloid-beta 42
(Abeta42), major components of amyloid plaques, and the cytotoxic C-terminal fragments,
gamma-CTF(50), gamma-CTF(57) and gamma-CTF(59). Many other minor beta-amyloid
peptides, beta-amyloid 1-X peptides, are found in cerebral spinal fluid (CSF) including the betaamyloid X-15 peptides, produced from the cleavage by alpha-secretase and all terminatiing at
Gln-686.
Proteolytically cleaved by caspases during neuronal apoptosis. Cleavage at Asp-739 by either
caspase-6, -8 or -9 results in the production of the neurotoxic C31 peptide and the increased
production of beta-amyloid peptides.
N- and O-glycosylated. O-glycosylation on Ser and Thr residues with core 1 or possibly core 8
glycans. Partial tyrosine glycosylation (Tyr-681) is found on some minor, short beta-amyloid
peptides (beta-amyloid 1-15, 1-16, 1-17, 1-18, 1-19 and 1-20) but not found on beta-amyloid 38,
beta-amyloid 40 nor on beta-amyloid 42. Modification on a tyrosine is unusual and is more
prevelant in AD patients. Glycans had Neu5AcHex(Neu5Ac)HexNAc-O-Tyr,
Neu5AcNeu5AcHex(Neu5Ac)HexNAc-O-Tyr and O-AcNeu5AcNeu5AcHex(Neu5Ac)HexNAc-OTyr structures, where O-Ac is O-acetylation of Neu5Ac. Neu5AcNeu5Ac is most likely Neu5Ac
2,8Neu5Ac linked. O-glycosylations in the vicinity of the cleavage sites may influence the
proteolytic processing. Appicans are L-APP isoforms with O-linked chondroitin sulfate.
Phosphorylation in the C-terminal on tyrosine, threonine and serine residues is neuron-specific.
Phosphorylation can affect APP processing, neuronal differentiation and interaction with other
proteins. Phosphorylated on Thr-743 in neuronal cells by Cdc5 kinase and Mapk10, in dividing
cells by Cdc2 kinase in a cell-cycle dependent manner with maximal levels at the G2/M phase
and, in vitro, by GSK-3-beta. The Thr-743 phosphorylated form causes a conformational change
which reduces binding of Fe65 family members. Phosphorylation on Tyr-757 is required for SHC
binding. Phosphorylated in the extracellular domain by casein kinases on both soluble and
membrane-bound APP. This phosphorylation is inhibited by heparin.
Extracellular binding and reduction of copper, results in a corresponding oxidation of Cys-144
and Cys-158, and the formation of a disulfide bond. In vitro, the APP-Cu(+) complex in the
presence of hydrogen peroxide results in an increased production of beta-amyloid-containing
peptides.
Trophic-factor deprivation triggers the cleavage of surface APP by beta-secretase to release
sAPP-beta which is further cleaved to release an N-terminal fragment of APP (N-APP).
Beta-amyloid peptides are degraded by IDE.
Cellular localization
Membrane. Membrane, clathrin-coated pit. Cell surface protein that rapidly becomes internalized
via clathrin-coated pits. During maturation, the immature APP (N-glycosylated in the
endoplasmic reticulum) moves to the Golgi complex where complete maturation occurs (O3
glycosylated and sulfated). After alpha-secretase cleavage, soluble APP is released into the
extracellular space and the C-terminal is internalized to endosomes and lysosomes. Some APP
accumulates in secretory transport vesicles leaving the late Golgi compartment and returns to the
cell surface. Gamma-CTF(59) peptide is located to both the cytoplasm and nuclei of neurons. It
can be translocated to the nucleus through association with APBB1 (Fe65). Beta-APP42
associates with FRPL1 at the cell surface and the complex is then rapidly internalized. APP
sorts to the basolateral surface in epithelial cells. During neuronal differentiation, the Thr-743
phosphorylated form is located mainly in growth cones, moderately in neurites and sparingly in
the cell body. Casein kinase phosphorylation can occur either at the cell surface or within a postGolgi compartment. Associates with GPC1 in perinuclear compartments. Colocalizes with
SORL1 in a vesicular pattern in cytoplasm and perinuclear regions.
Anti-Amyloid Precursor Protein antibody images
ab15272 at a 1/100 dilution staining Amyloid
Precursor protein from human HEK cells by
Immunocytochemistry. The antibody was
incubated with the cells for 12 hours and then
detected using a goat anti-rabbit IgG AlexaFluor ® 568.
The image is courtesy of an Abreview
submitted by Randal Moldrich on 25 January
2006.
Immunocytochemistry/ Immunofluorescence Amyloid Precursor Protein antibody (ab15272)
ab15272 at a 1/100 dilution staining Amyloid
Precursor Protein from Rat Cells
(Hippocampal neuron culture) by
Immunocytochemistry. The antibody was
incubated with the cells for 12 hours and then
detected using a goat anti-rabbit IgG AlexaFluor® 568.
The image is courtesy of an Abreview
submitted by Randal Moldrich on 25 January
Immunocytochemistry/ Immunofluorescence -
2006.
Amyloid Precursor Protein antibody (ab15272)
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Immunohistochemistry (Formalin/PFA-fixed
Immunohistochemistry (Formalin/PFA-fixed
paraffin-embedded sections) analysis of
paraffin-embedded sections) - Anti-Amyloid
human Alzheimers brain tissue labelling
Precursor Protein antibody (ab15272)
Amylois Precursor Protein with ab15272.
ab15272 staining Amyloid Precursor
Protein in 15 month old TASTPM mouse
brain tissue section by Immunohistochemistry
(Formalin/ PFA fixed paraffin-embedded
sections). Tissue underwent fixation in
formaldehyde, heat mediated antigen retrieval
in 10mM citric acid buffer (pH 6.0) and
blocking in 1% BSA for 10 minutes at 25°C.
Immunohistochemistry (Formalin/PFA-fixed
paraffin-embedded sections) - Amyloid Precursor
Protein antibody (ab15272)
This image is courtesy of an Abreview submitted by
Mr Carl Hobbs
The primary antibody was used
at 1/300 dilution, and incubated with
sample for 2 hours. A Biotin conjugated goat
polyclonal to rabbit IgG at 1/200 dilution, was
used as secondary.
ab15272 staining Amyloid Precursor Protein
in murine N2a cells by ICC
(Immunocytochemistry).
Cells were fixed with paraformaldehyde and
blocked with 10% serum for 15 minutes at
Immunocytochemistry - Anti-Amyloid Precursor
24°C. Samples were incubated with primary
Protein antibody (ab15272)
antibody (1/100 in 1% PBS) for 2 hours at
This image is courtesy of an anonymous Abreview
24°C. An biotin-conjugated goat anti-rabbit
polyclonal IgG (1/200) was used as the
secondary antibody.
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All lanes : Anti-Amyloid Precursor Protein
antibody (ab15272) at 1/1000 dilution
Lane 1 : Rat brain tissue lysate
Lane 2 : Rat brain tissue lysate
Lysates/proteins at 100 µg per lane.
Secondary
HRP-conjugated Goat anti-rabbit IgG
polyclonal at 1/2000 dilution
Western blot - Anti-Amyloid Precursor Protein
developed using the ECL technique
antibody (ab15272)
This image is courtesy of an anonymous Abreview
Performed under reducing conditions.
Predicted band size : 87 kDa
Observed band size : 87 kDa
Exposure time : 30 seconds
This image is courtesy of an anonymous
Abreview
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